Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/06—Dipeptides
  • C07K5/06008—Dipeptides with the first amino acid being neutral
  • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
  • C07K5/0606—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing heteroatoms not provided for by C07K5/06086 - C07K5/06139, e.g. Ser, Met, Cys, Thr
  • C07K5/06069—Ser-amino acid
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C323/00—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups
  • C07C323/50—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton
  • C07C323/51—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton
  • C07C323/60—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton with the carbon atom of at least one of the carboxyl groups bound to nitrogen atoms
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
  • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
  • Y02P20/00—Technologies relating to chemical industry
  • Y02P20/50—Improvements relating to the production of bulk chemicals
  • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Definitions

• the present invention relates to novel amides, a process for the preparation thereof, the pharmaceutically acceptable salts thereof and pharmaceutical compositions containing them.
• AA arachidonic acid
• AA Through cyclooxygenase, AA gives rise to prostaglandins and thromboxanes, the most significant being PGE, and TxA 2 , which participate directly in inflammation (Higgs et al. Annals of Clinical Research, 16, 287-299 (1984)).
• PGE Protaglandins
• TxA 2 TxA 2
• lipooxygenase AA produces leukotrienes, the most important being LTB 4 , LTC 4 and LTD 4 , which also participate in inflammatory reactions, showing chemotactic activities, stimulate segregation of lysosomal enzymes and act as important factors in the immediate hypersensitivity reactions (Bailey and Casey, Ann. Rep. Med. Chem. , 17, 203-217 (1982)).
• PLA 2 By the action of PLA 2 , besides the release of fatty acids, the corresponding lysophospholipids are obtained, which either can then be re-esterified or be converted into PAF (platelet activating factor) by acetylation at 2-position, if they have phosphocholine at the 3-position, an ether bond at the 1-position and are in a cell system having acetyl-transferase activity.
• PAF platelet activating factor
• PAF platelet activating factor
• PLA 2 besides being related to the inflammatory processes, can also be involved, either directly or indirectly, in degenerative thrombotic and cancerous pathologies.
• PLA 2 is of great importance in controlling the production of the mediators involved in the pathologies indicated above.
• compounds inhibiting PLA 2 provide a new rational approach for the prevention, elimination or improvement of different allergic, anaphylactic, asthmatic, inflammatory and thrombotic conditions.
• the compounds of general formula (I) are not structurally related to any PLA 2 inhibitor described in literature. Some compounds that could be considered structurally related to the compounds of the present invention are not included in formula (I) and show very different activities, for example some of them are receptor inhibitors of LTC 4 , (Sala et al., Eicosanoids, 3, 105 (1990)), or of glutathione S-transferase (Adang et al., J. Biol. Chem., 266, 830 (1991)), or they enhance the immune activity (JP 55085553); or they are S-butylglutathione-like metabolites (James et al., Bio-chem. J. 109, 727 (1968)).
• corticosteroids are the only medicaments considered likely to exert an inhibitory mechanism on PLA 2 , even though in an indirect way.
• said compounds have a series of adverse systemic side-effects which restrict the use thereof, particularly in chronic pathologies.
• PLA 2 selective inhibitors, such as the compounds of the present invention advantageously have the same effectiveness as corticosteroids, without the side-effects thereof.
• - X is an oxygen or sulphur atom
• - R 1 is a C 4 -C 20 straight or branched alkyl group ;
• R 2 is hydrogen or a R 5 -CO-, R 5 -O-CO- or R 5 -SO 2 -group, in which R 5 is a C 1 -C 20 straight or branched alkyl group, phenyl or an arylalkyl group of less than 20 carbon atoms totally;
• R 3 is hydrogen, a carboxy group, an alkoxycarbonyl, aryloxycarbonyl or arylalkoxycarbonyl group of less than 10 carbon atoms in the last three cases;
• R 4 is hydrogen, a C 1 -C 6 straight or branched alkyl group, an arylalkyl group of less than 10 carbon atoms, an heteroarylalkyl group of less than 10 carbon atoms, an hydroxyalkyl group of less than 4 carbon atoms, a thioalkyl or alkylthioalkyl group of less than 4 carbon atoms, an aminoalkyl group of less than 6 carbon atoms, the amino group being in the free or derivatized form, as an alkyl amide of less than 4 carbon atoms, a C 2 -C 5 carboxyalkyl group, the carboxy group being in the free or derivatized form as an alkyl or aralkyl ester of less than 8 carbon atoms, a carbamoylalkyl group of less than 4 carbon atoms, a guanidinoalkyl group of less than 5 carbon atoms, or a sulfoalkyl group of less than 4 carbon
• X- is a pharmaceutically acceptable anion of an inorganic acid (e.g. hydrochloric or sulfuric) or organic carboxylic acid (e.g. acetic, trifluoroacetic, lactic or tartaric), or organic sulfonic acid (e.g. methanesulfonic, ethanesulfonic or toluenesulfonic).
• inorganic acid e.g. hydrochloric or sulfuric
• organic carboxylic acid e.g. acetic, trifluoroacetic, lactic or tartaric
• organic sulfonic acid e.g. methanesulfonic, ethanesulfonic or toluenesulfonic.
• M + is an alkali metal cation (e.g. Na + , K + ), or the equivalent of an alkaline-earth metal cation (e.g. 1/2 Ca 2 +, 1/2 Mg 2+ ).
• the double acid addition salts can also be obtained (e.g. dihydrochlorides or dihydrobromides).
• the double base addition salts can also be obtained (e.g. sodium, potassium, calcium or magnesium salts).
• the compounds of general formula (I) have one or more asymmetric carbons in their structure.
• the present invention includes all the possible stereoisomers as well as the mixtures thereof.
• R 1 can be for example butyl, hexyl, decyl, tetradecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl or icosyl; when R 2 is a R 5 -CO-, R 5 -O-CO- or R -SO 2 -, in which R 5 is an alkyl or arylalkyl group, this can be methyl, ethyl, propyl, butyl, decyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, icosyl, benzyl or 11-phenylundecyl; when R 3 is an alkoxycarbonyl, aryloxycarbonyl or arylalkoxycarbonyl group, they can be methoxycarbonyl, ethoxycarbonyl
• Preferred compounds of the present invention are those in which:
• R 1 are the groups defined above;
• - R 2 is hydrogen, a C 1 -C 21 alkoxycarbonyl group atoms, preferably groups acetyl, hexanoyl, dodecanoyl or hexadecanoyl;
• - R 3 is hydrogen, a carboxy group or an alkoxycarbonyl group of less than 10 carbon atoms, preferably methoxycarbonyl or ethoxycarbonyl;
• R 4 is hydrogen, a C 1 -C 6 straight or branched alkyl group, preferably methyl or isobutyl, an hydroxyalkyl group of less than 4 carbon atoms, preferably hydroxymethyl or 2-hydroxyethyl, an aminoalkyl or alkylcarbonylaminoalkyl group of less than 7 carbon atoms totally, preferably 4-aminobutyl or 4-acetylaminobutyl, a C 2 -C 5 carboxyalkyl group, preferably carboxymethyl or
• 2-carboxyethyl or a sulfoalkyl of less than 4 carbon atoms, preferably sulfomethyl.
• Particularly preferred compounds of the present invention are the following:
• N-Acetyl-S-decyl-D ,L-cysteinyltaurine ( sodium salt ) N-Acetyl-S-decyl-D ,L-cysteinyltaurine ( sodium salt ) .
• N-Acetyl-S-octadecyl-D-cysteinyltaurine sodium salt
• the compounds of general formula ( I ) can be prepared by reacting a compound of formula ( II) , of suitable stereochemistry at the carbon at the 2-position
• R 6 is a suitable aminoprotecting group, for example benzyloxycarbonyl or tert-butyloxycarbonyl, with:
• R 3 is one of the groups defined above, except for hydrogen
• R 4 is one of the groups defined above, except for a sulfoalkyl group, a reactive of formula (III), of suitable stereochemistry
• R 7 can be equivalent to R 3 in formula (I), except for hydrogen, and, when in (I) R is a carboxy group, then R 7 is a suitably protected carboxy group, for example in form of the benzyl or tert-butyl ester, and R 8 can be the same as R 4 , except when in formula (I) R 4 contains a functional group in the free form, in which case in R 8 such a group is suitably protected, for example when R 4 in formula (I) contains an amino group, R 8 in formula (III) can contain a benzyloxycarbonylamino or tert-butyloxycarbonylamino group; when R 4 in (I) contains a carboxy group, this is for example in form of the benzyl or tert-butyl ester in (III); the reaction between (II) and (III) is carried out in the presence of a proton-binding base, such as triethylamine or pyridine and a carboxy-activating agent,
• This compound of formula ( IV) is converted into ( I ) by removing any protecting groups present in R 6 , R 7 g and R 8 , according to conventional methods; when R 6 and R 7 are benzyloxycarbonyl groups and a benzyloxycarbonylamino or a benzyloxycarbonyl group can be present in R 8 , the benzyl groups can be removed by catalytic hydrogenation with Pd-C or with Pd(OH) 2 , in solvents such as methanol, water or acetic acid, under hydrogen pressures from atmospheric pressure to 50 psi, at a tempe rature from 20° to 50°C for a time from 3 to 20 hours; or in an acid medium, for example with hydrobromic or trifluoroacetic acid, in a suitable solvent such as acetic acid or chloroform, at a temperature from 0o to
• R 6 and R 7 are tert-butyloxycarbonyl groups and a tert-butyloxycarbonylamino or tert-butyloxycarbonyl group can be present in R 8 , the tert-butyl groups can be removed under the same acid conditions as mentioned above .
• R 6 in formula ( IV) is an amino-protecting group , such as benzyloxycarbonyl or tert-butyloxycarbonyl , this is deprotected to obtain compounds of formula ( I) wherein R 2 is hydrogen , which can in their turn be converted into the compounds (I. with R 2 different from hydrogen by an acylation or sulfonylation reaction with suitable acid halides or anhydrides (R 5 COX, R 5 OCOX or R 5 SO 2 X).
• N-hydroxy succinimide in the presence of a dehydrating agent such as dicyclohexylcarbodiimide, in a suitable solvent such as dioxane, tetrahydrofuran or N,N-dimethylformamide, at a temperature from 0o to 40°C for a time from 5 to 24 hours. Subsequently, it is reacted with a reactive (V)
• R 4 is a sulfoalkyl group of less than 4 carbon atoms, in the presence of a base such as sodium bicarbonate or aqueous sodium hydroxide, in a suitable organic solvent such as dioxane or tetrahydrofuran at a temperature from 0oC to the solvent's reflux, for a time from 5 to 24 hours, to obtain a compound of formula (VI)
• this compound (VI) coincides with compound (I) wherein R 3 is hydrogen and with the proviso that R 2 is hydrogen.
• R 6 is an amino-protecting group
• this compound can be transformed into compound (I. wherein R 2 is hydrogen, by means of con- ventional deprotection methods, or optionally it can be converted into the remaining compounds (I), as indicated above.
• a starting compound (II) wherein X is sulphur can be prepared, for example, following the synthetic sequence shown in Scheme 1.
• a compound of formula (VIII) wherein R 6 is one of the groups defined above can be obtained (step a) starting from cysteine ((VII)) of suitable stereochemistry, this product being commercially available in both the chiral D or L forms and the racemic one, by reaction with a suitable acyl or sulfonyl halide R 6 Y, wherein R 6 is the group described above and Y can be iodide, chloride or bromide, in the presence of a base such as trie thy lamine, sodium acetate or sodium bicarbonate, in a suitable solvent such as water or N,N-dimethylformamide, at a temperature from 0oC to the solvent's reflux, for a time from 1 to 12 hours.
• a suitable acyl or sulfonyl halide R 6 Y wherein R 6 is the group described above and Y can be iodide, chloride or bromide, in the presence of a base such as trie thy la
• a compound of formula (II) can be obtained (step b) by reacting the compound (VIII) with a compound ZR , wherein R 1 is the group described above and Z is a suitable leaving group, such as for example bromide, iodide, mesylate or tosylate in the presence of a base such as potassium hydroxide or sodium carbonate, in a suitable organic solvent such as ethanol or N,N-dimethylformamide, at a temperature from 0° to 50°C, for a time from 5 to 24 hours.
• a base such as potassium hydroxide or sodium carbonate
• a suitable organic solvent such as ethanol or N,N-dimethylformamide
• a starting compound (II) wherein X is oxygen can be prepared, for example, following the synthetic sequence shown in Scheme 2.
• a compound of formula (X) can be prepared, for example, starting from compound (IX), the preparation of which is well described in literature (Hajdu et al.
• a compound of formula (XI), wherein R 1 and R 6 are the groups defined above, can be obtained (step d), by means of reaction with a suitable acyl or sulfonyl halide as described above.
• a compound of formula (II) wherein X is oxygen can be prepared, for example, by subjecting a compound (XI) to the action of an oxidizing agent such as the system formed by RuCl 3 .3H 2 O/NaIO 4 , in mixtures of suitable solvents such as carbon tetrachloride:acetonitrile:water (3:3:2) or carbon tetrachloride:ethyl acetate:water (3:3:2), at a temperature from -20° to 30oC, for a time from 15 min to 18 hours.
• an oxidizing agent such as the system formed by RuCl 3 .3H 2 O/NaIO 4
• suitable solvents such as carbon tetrachloride:acetonitrile:water (3:3:2) or carbon tetrachloride:ethyl acetate:water (3:3:2)
• the compounds of the present invention show a marked activity as inhibitors of PLA 2 activity, and accordingly they have antiinflammatory and antiallergic properties which make them useful for the treatment of diseases in which this enzyme is involved.
• said compounds can be used in human therapy for the prevention and treatment of allergic rhinitis, bronchial asthma, hypersensitivity reactions such as allergic conjunctivitis, various inflammatory conditions such as those present in rheumatoid arthritis, osteoarthritis, tendinitis, bursitis, psoriasis and other related inflammations.
• the compounds of the present invention are formulated in the appropriate pharmaceutical form, according to conventional techniques and excipients, such as those described in Remington's Pharmaceutical Science Handbook, Mack Pub. Co., N.Y., USA.
• Examples of said forms include capsules, tablets, syrups and the like, containing from 1 to 1000 mg per unitary dose.
• N.M.R. 1 H (300 MHz, CDCl 3 ) ⁇ ppm: 0.85 (t, 3H); 1.25 (m, 30H) ; 1.51 (m, 2H) ; 2 .49 (t , 2H ) ; 2 . 98 ( d, 2H) ; 4 . 58 ( m, 1H) ; 5. 10 ( s , 2H ) ; 7 .30 (m, 5H ) .
• N.M.R. 1 H (300 MHz, CDCl 3 ) ⁇ ppm: 0.85 (t, 3H); 1.32 (m, 2H); 1.50 (m, 2H); 2.49 (t, 2H); 2.98 (d, 2H); 4.58 (m, 1H); 5.10 (s, 2H); 7.30 (m, 5H).
• R 7 COOCH 2 C 6 H 5
• R 8 CH 2 CH (CH 3 ) 2 ).
• N.M.R. 1 H (300 MHz, CDCl 3 ) ⁇ ppm: 0.85 (t, 9H); 1.24 (m, 30H); 1.56 (m, 5H); 2.50 (m, 2H); 2.79 (dd, 1H);
• R 6 and R 7 COOCH 2 C 6 H 5
• R 8 (CH 2 ) 4 NHCOOCH 2 C 6 H 5 ).
• R 6 and R 7 COOCH 2 C 6 H 5
• R 8 (CH 2 ) 4 NHCOOCH 2 C 6 H 5 ).
• N.M.R. 1 H (300 MHz, CDCl 3 ) S ppm: 0.86 (t, 3H); 1.25 (m, 30H); 1.40-1.90 (complex signal, 8H); 2.48 (m, 2H);
• N.M.R. 1 H (300 MHz, CDCl 3 ) ⁇ ppm: 0.86 (t, 3H); 1.25 (m, 30H); 1.40-1.90 (complex signal, 8H); 2.48 (m, 2H); 2.84 (m, 2H); 3.05 (m, 2H); 4.30 (m, 1H); 4.58 (m, 1H); 5.10 (m, 6H); 7.30 (m, 15H).
• N.M.R. 1 H (300 MHz, CDCl 3 ) ⁇ ppm: 0.86 (t, 3H); 1.25 (m, 30H); 1.40-1.90 (complex signal, 8H); 2.48 (m, 2H); 2.84 (m, 2H); 3.05 (m, 2H); 4.30 (m, 1H); 4.58 (m, 1H); 5.10 (m, 6H); 7.30 (m, 15H).
• N.M.R. 1 H (300 MHz, CDCl 3 ) ⁇ ppm: 0.88 (t, 3H); 1.35 (m, 2H); 1.50 (m, 2H); 2.02 (m, 1H); 2.24 (m, 1H); 2.40 (m, 2H); 2.50 (m, 2H); 2.88 (m, 2H); 4.39 (m, 1H); 4.69 (m, 1H); 5.10 (m, 6H); 7.26 (m, 15H).
• N.M.R. 1 H (300 MHz, CDCl 3 ) ⁇ ppm: 0.85 (t, 3H); 1.20-1.90 (complex signal, 10H); 2.48 (t, 2H); 2.70 (dd, 1H); 2.88 (dd, 1H); 3.08 (m, 2H); 4.30 (m, 1H); 4.59 (m, 1H); 5.10 (m, 6H); 7.30 (m, 15H).
• N.M.R. 1 H (300 MHz, CD 3 OD) ⁇ ppm: 0.89 (t, 3H); 1.29 (m, 30H); 1.60 (m, 2H); 2.02 (m, 1H); 2.22 (m, 1H); 2.43 (m, 2H); 2.62 (m, 2H); 3.00 (m, 2H); 4.10 (m, 1H); 4.54 (m, 1H); 5.19 (m, 2H); 7.34 (m, 5H).
• N.M.R. 1 H (300 MHz, CDCl 3 ) ⁇ ppm: 0.85 (m, 6H); 1.20 (m, 30H); 1.56 (m, 5H); 2.58 (m, 2H); 3.15 (m, 2H); 3.74 (m, 1H); 4.50 (m, 1H); 5.28 (q, 2H); 7.30 (m, 5H) .
• N.M.R. 1 H (300 MHZ, CD 3 OD) ⁇ ppm: 0.90 (t, 3H); 1.30 (m, 30H); 1.49-2.02 (complex signal, 8H); 2.65 (t, 2H);
• N.M.R. 1 H (300 MHz, CD 3 OD) ⁇ ppm: 0.90 (t, 3H); 1.30 (m, 30H); 1.49-2.02 (complex signal, 8H); 2.65 (t, 2H);
• R 4 CH 2 CH(CH 3 ) 2 ).
• N.M.R. 1 H (300 MHz, CD 3 OD) ⁇ ppm: 0.94 (t, 3H); 1.39-2.00 (complex signal, 10H); 2.67 (t, 2H); 2.96 (m, 3H); 3.14 (dd, 1H); 4.19 (t, 1H); 4.43 (m, 1H).
• R 4 CH 2 CH 2 COOH) .
• N.M.R. 1 H (300 MHz, CD 3 OD) ⁇ Ppm: 0.92 (t, 3H); 1.29 (m, 30H); 1.58 (m, 2H); 1.97 (m, 1H); 2.00 (s, 3H);
• R 4 CH 2 CH(CH 3 ) 2 ).
• N.M.R. 1 ⁇ (300 MHz, CD 3 OD) ⁇ ppm: 0.89 (t, 3H); 1.33 (m, 30H); 1.54 (m, 2H); 2.01 (s, 3H); 2.55 (t, 2H);
• R 4 (CH 2 ) 4 NHCOCH 3 ).
• N.M.R. 1 H (300 MHz, CD 3 OD) ⁇ ppm: 0.90 (t, 3H); 1.29 (m, 30H); 1.34-1.64 (complex signal, 6H); 1.73 (m, 1H); 1.89 (m, 1H); 1.92 (s, 3H); 2.02 (s, 3H); 2.56 (t, 2H);
• R 4 (CH 2 ) 4 NHCOCH 3 ).
• N.M.R. 1 H (300 MHz, CD 3 OD) ⁇ ppm: 0.90 (t, 3H); 1.29 (m, 30H); 1.34-1.64 (complex signal, 6H); 1.73 (m, 1H);
• R 4 (CH 2 ) 4 NHCOCH 3 ).
• N.M.R. 1 H (300 MHz, CD 3 OD) ⁇ ppm: 0.90 (t, 3H); 1.29 (m, 30H); 1.34-1.64 (complex signal, 6H); 1.73 (m, 1H);
• R 4 (CH 2 ) 4 NHCOCH 3 ).
• R 4 CH 2 CH 2 COOH) .
• N.M.R. 1 H (300 MHz, CD 3 OD) ⁇ ppm: 0.90 (m, 6H); 1.22- 1.46 (complex signal, 18H); 1.56 (m, 4H); 1.96 (m, 1H); 2.18 (m, 1H); 2.25 (t, 2H); 2.40 (q, 2H); 2.55 (t, 2H); 2.73 (m, 1H); 2.93 (m, 1H); 4.44 (m, 1H); 4.56 (m, 1H).
• N.M.R. 1 H (300 MHz, CD 3 OD) ⁇ ppm: 0.92 (t, 3H); 1.39-1.64 (complex signal, 8H); 1.74 (m, 1H); 1.86 (m, 1H); 1.91 (s, 3H); 1.99 (s, 3H); 2.56 (t, 2H); 2.72 (dd, 1H); 2.92 (dd, 1H); 3.14 (t, 2H); 4.38 (m, 1H); 4.56 (t, 1H).
• N.M.R. 1 H (300 MHz, CD 3 OD) ⁇ ppm: 0.92 (t, 3H); 1.34 (m, 30H); 1.65 (m, 2H); 2.65 (t, 2H); 3.04 (dd, 1H); 3.18 (dd, 1H); 4.22 (dd, 1H).
• N.M.R. 1 H (300 MHz, CDCl 3 ) ⁇ ppm: 0.85 (t, 3H); 1.25 (m, 30H); 1.55 (m, 2H); 2.06 (s, 3H); 2.50 (t, 2H); 2.99 (m, 2H); 4.72 (m, 1H).
• N.M.R. 1 H (300 MHz, CD 3 0D) ⁇ ppm: 0.89 (t, 3H); 1.29 (m, 30H); 1.58 (m, 2H); 1.92 (s, 3H); 2.55 (t, 2H); 2.71 (m, 1H); 2.96 (m, 3H); 3.59 (m, 2H); 4.44 (m, 1H).
• N.M.R. 1 H (300 MHz, CDCl 3 ) ⁇ ppm: 0.85 (t, 3H); 1.24 (m, 30H); 1.55 (m, 2H); 2.00 (s, 3H); 3.40 (t, 2H); 3.50-3.68 (complex signal, 3H); 3.80 (dd, 1H); 4.03 (m,
• N.M.R. 1 H (300 MHz, CDCl 3 ) ⁇ ppm: 0.85 (t, 3H); 1.25 (m, 30H); 1.50 (m, 2H); 1.99(m, 1H); 2.00 (s, 3H); 2.21 (m, 1H); 2.38 (m,2H); 3.39 (m, 3H); 3.76 (dd, 1H); 4.51 (m, 1H); 4.63 (m, 1H); 5.08 (s, 2H); 5.12 (s, 2H); 7.32 (m, 10H).
• R 8 CH 2 CH(CH 3 ) 2 ). According to the procedure described in example 6, starting from N-acetyl-O-octadecyl-D,L-serine and benzyl L-leucinate (hydrochloride), the title compound was prepared as a colourless oil (92 % yield).
• N.M.R. 1 H (300 MHz, CDCl 3 ) ⁇ ppm: 0.87 (m, 9H); 1.21 (m, 30H); 1.61 (m, 5H); 1.98 (s, 3H); 3.42 (m, 3H); 3.78 (m, 1H); 4.50 (m, 1H); 4.62 (m, 1H); 5.12 (s, 2H); 7.32 (m, 5H).
• N.M.R. 300 MHz, CD 3 OD
• 0.90 0.90 (t, 3H); 1.28 (m, 30H); 1.56 (m, 2H); 1.95 (m, 1H); 2.02 (s, 3H); 2.20 (m, 1H); 2.41 (m,2H); 3.46 (m, 2H); 3.65 (m, 2H); 4.46 (m, 1H); 4.58 (m, 1H).
• N.M.R. 1 H (300 MHz, CD 3 OD) ⁇ ppm: 0.92 (m, 9H); 1.29 (m, 30H); 1.49-1.74 (complex signal, 5H); 2.00 (s, 3H); 3.46 (m, 2H); 3.65 (m, 2H); 4.45 (m, 1H); 4.58 (m, 1H).
• the PLA 2 activity is determined by radioactive evaluation of the labelled fatty acid which esterifies the sn-2 position of the phospholipid substrate and which is released by the enzyme action.
• the used enzyme is purified PLA 2 from human synovial liquid.
• the reaction mixture contains 25 mM Hepes buffer (pH 7.0), 5.0 mM Ca 2+ and 1.4 ⁇ 106 E. coli autoclaved (corresponding to 10000 dpm and 10.0 nmol of phospholipid).
• the reaction is started by addition of 80 ng of purified enzyme, keeping stirring for 5 minutes at 37oC
• the reaction is stopped by adding 3.0 ml of CHCl 3 :CH 3 OH (1:2 v/v), the lipids are extracted by the Bligh and Dyer procedure and the reaction products are separated by thin layer chromatography and dpm are quantified by liquid scintillation.
• the compounds of the present invention were tested according to this procedure.
• the inhibiting action of the different compounds is expressed as the product concentration inhibiting the activity of the enzyme by 50% compared with the control in the absence of inhibitor (IC 50 ).
• Some examples of the obtained results are reported in the following Table.

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